Abstract
BackgroundMiRNAs are a group of multifunctional non-coding RNAs which play an important role in the various physiological processes including the development of NAFLD. Recent studies have shown that miR-30b-5p tightly associated with the abnormal lipid metabolism in patients with NAFLD, but the detailed mechanism of miR-30b-5p in the lipid metabolism was remain unclear. The aim of this study was to investigate the effect of miR-30b-5p on the lipid metabolism in hepatocellular carcinoma Huh-7 cells.Material and methodsThe correlation of intracellular fat content with the expression of miR-30b-5p in Huh-7 cells and HepG2 cells was investigated by treated cells with different concentrations of FFAs. The effect of miR-30b-5p on the lipid deposition in Huh-7 cells was tested by oil red O staining and TG concentrations measurement. qRT-PCR and western blot were used to investigate the lipid metabolism-related genes PPAR-α, SREBP-1, and GULT1 in miR-30b-5p overexpressed or inhibited Huh-7 cells. Target genes of miR-30b-5p were predicted using starBase, miRDB, and TargetScan databases and verified by qRT-PCR and western blot.ResultsThe expression of miR-30b-5p was significant decreased in the FFAs treated Huh-7 cells and HepG2 cells. Overexpressing miR-30b-5p in Huh-7 cells decreased the number and size of lipid droplets and intracellular TG concentrations in Huh-7 cells. Expression of fatty acid oxidation related gene PPAR-α was increased and expression of lipid synthesis related gene SREBP-1 was decreased in the miR-30b-5p overexpressed Huh-7 cells. In addition, miR-30b-5p regulates the intracellular lipid metabolism by targeting PPARGC1A.ConclusionsOverexpression of miR-30b-5p could reduce the intracellular fat deposition in Huh-7 cells, and miR-30b-5p might regulate the intracellular lipid metabolism by targeting the PPARGC1A in Huh-7 cells.
Highlights
Non-alcoholic fatty liver disease (NAFLD) is caused by the excessive lipid accumulation in the liver and is thought to be the manifestation of metabolic syndrome in the liver [1]
Expression of miR-30b-5p negative correlated with lipid content in Hun-7 cells and HepG2 cells To investigate the effect of lipid overload on the expression of miR-30b-5p in Huh-7 cells and HepG2 cells, we treated the Huh-7 cells and HepG2 cells with 0.5 mM and 1 mM Free fatty acid (FFA) for 24 h
These results proved that lipid overload in Huh-7 cells and HepG2 cells could inhibit the expression of miR30b-5p, suggested that miR-30b-5p may play an important role in the lipid metabolism of Huh-7 cells and HepG2 cells
Summary
Non-alcoholic fatty liver disease (NAFLD) is caused by the excessive lipid accumulation in the liver and is thought to be the manifestation of metabolic syndrome in the liver [1]. Extracellular vesicles can be absorbed by other cells through receptormediated or receptor-independent ways and release the miRNAs to regulate the gene expressions in the cells [16, 17]. MiRNAs play an important role in tumor development and metabolic diseases, and they possess significant potential and application value in early disease detection, treatment, and monitoring disease progression, as well as organism’s response to various treatments [18]. MiRNAs are a group of multifunctional non-coding RNAs which play an important role in the various physiological processes including the development of NAFLD. The aim of this study was to investigate the effect of miR-30b-5p on the lipid metabolism in hepatocellular carcinoma Huh-7 cells
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